Advances, Perspectives and Potential Engineering Strategies of Light-Gated Phosphodiesterases for Optogenetic Applications

Abstract

The second messengers, cyclic adenosine 3′-5′-monophosphate (cAMP) and cyclic guanosine 3′-5′-monophosphate (cGMP), play important roles in many animal cells by regulating intracellular signaling pathways and modulating cell physiology. Environmental cues like temperature, light, and chemical compounds can stimulate cell surface receptors and trigger the generation of second messengers and the following regulations. The spread of cAMP and cGMP is further shaped by cyclic nucleotide phosphodiesterases (PDEs) for orchestration of intracellular microdomain signaling. However, localized intracellular cAMP and cGMP signaling requires further investigation. Optogenetic manipulation of cAMP and cGMP offers new opportunities for spatio-temporally precise study of their signaling mechanism. Light-gated nucleotide cyclases are well developed and applied for cAMP/cGMP manipulation. Recently discovered rhodopsin phosphodiesterase genes from protists established a new and direct biological connection between light and PDEs. Light-regulated PDEs are under development, and of demand to complete the toolkit for cAMP/cGMP manipulation. In this review, we summarize the state of the art, pros and cons of artificial and natural light-regulated PDEs, and discuss potential new strategies of developing light-gated PDEs for optogenetic manipulation.

Characterization and Modification of Light-Sensitive Phosphodiesterases from Choanoflagellates

“…However, the current light-regulated PDEs need development, as strong light-regulation of PDE activity is still missing [ 29 ]. To this purpose, light-activated phosphodiesterase (LAPD) was first engineered by combining a bacterial photosensor module with the catalytic domain of human PDE2A [ 30 , 31 ].…”

“…Sr RhoPDE was the first natural photoreceptor discovered with light-enhanced phosphodiesterase activity [ 14 , 15 ]. Current available light-activated PDEs, artificial LAPD and natural RhoPDEs, showed similar drawbacks of relatively high dark activity and low light to dark activity ratio [ 29 ]. Therefore, searching and engineering of new light-gated RhoPDEs will provide more possibilities for optogenetic cGMP/cAMP manipulation.…”

Photoactivated adenylyl cyclases attenuate sepsis-induced cardiomyopathy by suppressing macrophage-mediated inflammation

“…But it seems that bPAC or biPAC may modulate the expression of IL-1 and TNF-a through diverse signaling pathways, which is certainly deserved to be further e xplor ed. Fu rt her mor e, the l ight-ac tiv ate d c A MP phosphodiesterases (29,30) can be applied together with PACs for bidirectional regulation of immune cells. Actually, we are working on constructing subcellular optogenetics molecules through anchor proteins such as AKAP79, as the compartment of AC expression also has substantial consequence on its biological function.…”

From the Sea for the Sight: Marine Derived Products for Human Vision

“…Examples are the BeCyclop (also known as RhGC) identified in the aquatic fungus Blastocladiella emersonii (Avelar et al, 2014 ; Gao et al, 2015 ; Scheib et al, 2015 ) and the SrRhoPDE from a marine species of protists, the Choanoflagellate Salpingoeca rosetta , proven as blue light-activated phosphodiesterase, degrading cGMP and cAMP (Lamarche et al, 2017 ; Yoshida et al, 2017 ). The properties of these last opsins have stimulated strong interest in order to expand the use of light stimuli for controlling intracellular signaling and specific biochemical events in cells, also by engineering non-opsin proteins able to modulate general second messengers (Tian et al, 2020 ).…”